UCHL1 attenuates diabetic retinopathy by deubiquitinating and stabilizing NRF2 to suppress oxidative stress-induced retinal vascular endothelial ferroptosis.

Abstract

Diabetic retinopathy (DR) is characterized by progressive retinal microvascular injury, with oxidative stress and ferroptosis increasingly recognized as key pathogenic contributors. This study investigated whether ubiquitin C-terminal hydrolase L1 (UCHL1) regulates retinal endothelial ferroptosis through stabilization of nuclear factor erythroid 2-related factor 2 (NRF2). Analysis of the GSE102485 dataset identified UCHL1 as a downregulated deubiquitinating enzyme in DR. Streptozotocin-induced type 1 diabetic mice and high glucose (HG)-induced human retinal capillary endothelial cells (HRCECs) were used, together with UCHL1 inhibition/knockdown, AAV-mediated UCHL1 overexpression, and NRF2 knockdown. UCHL1 expression was reduced in diabetic retinas and HG-induced HRCECs, accompanied by ferroptosis activation, mitochondrial injury, and endothelial dysfunction. Pharmacological inhibition or siRNA-mediated depletion of UCHL1 intensified oxidative stress, Fe2⁺ accumulation, lipid peroxidation, mitochondrial fragmentation, and cristae disruption, while impairing endothelial barrier integrity, migration, and tube formation. In vivo, LDN57444 aggravated retinal vascular leakage and fundus vascular abnormalities, whereas AAV-UCHL1 preserved retinal architecture and reduced vascular permeability. Mechanistically, co-immunoprecipitation and ubiquitination assays demonstrated that UCHL1 interacted with NRF2 and stabilized NRF2 by removing K48-linked polyubiquitin chains. Nuclear-cytoplasmic fractionation further showed that UCHL1 overexpression restored NRF2 abundance and increased nuclear NRF2 accumulation under HG conditions. Ferrostatin-1 rescued UCHL1 depletion-induced ferroptotic injury, whereas NRF2 knockdown abolished the protection conferred by UCHL1 overexpression. These findings highlight the UCHL1/NRF2 axis may represent a therapeutic target in DR.

Key Findings

• UCHL1 expression is reduced in diabetic retinas and high glucose-induced human retinal capillary endothelial cells, correlating with ferroptosis activation and endothelial dysfunction.
• UCHL1 stabilizes NRF2 by deubiquitinating and removing K48-linked polyubiquitin chains, enhancing NRF2 nuclear accumulation under high glucose conditions.
• Manipulating UCHL1 levels affects retinal vascular integrity and ferroptotic injury, with UCHL1 overexpression protecting against retinal vascular damage and ferroptosis in diabetic retinopathy models.

Clinical Significance

Citation

Xu Sanhua, Huang Jun, Wang Yicanget al.. UCHL1 attenuates diabetic retinopathy by deubiquitinating and stabilizing NRF2 to suppress oxidative stress-induced retinal vascular endothelial ferroptosis. Biochemical pharmacology. 2026-Aug-21.